The present technology relates to a method for fabricating a microchip for nucleic acid amplification reaction. More specifically, the present technology relates to a microchip for nucleic acid amplification reaction in which a solidified reagent that includes at least one or more kinds of the substances required for a reaction is contained in a well that serves as a reaction site for a nucleic acid amplification reaction.
In recent years, microchips have been developed in which wells and channels for performing chemical and biological analyses are provided on a silicon substrate or a glass substrate by applying micro-machining techniques used in the semiconductor industry. These microchips have begun to be utilized for electrochemical detectors in, for example, liquid chromatography, compact electrochemical sensors in medical service locations and the like.
Analytical systems using such microchips are called μ-TAS (micro-Total-Analysis System), lab-on-a-chip, bio chip or the like. Attention is being paid to such microchips as a technology that enables chemical and biological analyses to be performed faster, with greater efficiency, and a higher level of integration, or that enables the analyzing apparatuses to be reduced in size. μ-TAS, which enables analysis with a small amount of sample and enables the disposable use of microchips, is expected to be applied particularly in biological analyses where precious trace amounts of samples or many specimens are handled.
An applied example of μ-TAS is an optical detection apparatus in which a substance is introduced into a plurality of areas arranged on the microchip, and the substance is optically detected. Such an optical detection apparatus may include a reaction apparatus (for example, a real-time PCR apparatus) that causes a reaction, such as a nucleic acid amplification reaction, between a plurality of substances to proceed in a well on the microchip, and optically detects the produced substances.
Microchip-type nucleic acid amplification apparatuses have conventionally employed a method in which the reaction is performed by mixing in advance all of the reagents and template DNA required for the nucleic acid amplification reaction, and introducing the mixed solution into a plurality of wells arranged on the microchip. However, with this method, since it takes a certain amount of time until the mixed solution is introduced into the wells, there is the problem that during that period the reaction proceeds in the mixed liquid, so that non-specific nucleic acid amplification tends to occur, thereby reducing quantitative performance.
In response to the above problem, for example, JP-A-2011-160728 discloses a microchip in which a plurality of reagents required for a nucleic acid amplification reaction are laminated and fixed in order in the wells.